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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1
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pubmed:dateCreated |
1984-5-11
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pubmed:abstractText |
Polysomes containing cytosolic malic enzyme mRNA and malic enzyme nascent chains were complexed with specific antibodies and purified by chromatography on protein A-Sepharose. When poly(A+) mRNA derived from the immunoselected polysomes was translated in vitro, full length malic enzyme (subunit Mr = 58,000) accounted for a significant fraction (approximately 20%) of the polypeptides synthesized. Double-stranded cDNA, synthesized using partially purified malic enzyme mRNA as a template, was inserted into pBR 322 and cloned. Twenty-five candidate malic enzyme cDNA clones were identified by differential hybridization. Four clones were studied further and each of these was shown to have malic enzyme cDNA sequences by hybrid-selected translation and specific immunoprecipitation. Plasmid pME1, which contains a 1400-base pair insert, hybridized to two mouse liver malic enzyme mRNAs with lengths of 2300 and 3500 bases. Similar analyses were performed on liver mRNAs isolated from MOD-1 mutant mice which lack cytosolic malic enzyme activity. These Northern blots disclosed a pair of aberrantly large malic enzyme mRNAs with lengths of 2800 and 4000 bases. Furthermore, anti-malic enzyme antibodies exclusively precipitated a polypeptide translation product with a Mr of 77,000 when MOD-1 mRNA was used to direct in vitro protein synthesis. Thus, it is possible that MOD-1 malic enzyme mRNA contains an additional polypeptide coding sequence. The translation of such a sequence might disrupt enzyme function and/or markedly decrease enzyme stability. The malic enzyme cDNA probe was also employed to demonstrate that the induction of malic enzyme in the livers of previously starved mice that were fed a high carbohydrate, fat-free diet was controlled pretranslationally by a parallel modulation of the malic enzyme mRNA concentration.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
10
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pubmed:volume |
259
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
555-9
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:6546753-Animals,
pubmed-meshheading:6546753-Base Sequence,
pubmed-meshheading:6546753-Cloning, Molecular,
pubmed-meshheading:6546753-DNA,
pubmed-meshheading:6546753-Liver,
pubmed-meshheading:6546753-Malate Dehydrogenase,
pubmed-meshheading:6546753-Mice,
pubmed-meshheading:6546753-Mice, Mutant Strains,
pubmed-meshheading:6546753-Nucleic Acid Hybridization,
pubmed-meshheading:6546753-RNA, Messenger
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pubmed:year |
1984
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pubmed:articleTitle |
Cloning of cDNA sequences for murine malic enzyme and the identification of aberrantly large malic enzyme mRNA in MOD-1 null mice.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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